Document Detail

Structure of an argonaute silencing complex with a seed-containing guide DNA and target RNA duplex.
MedLine Citation:
PMID:  19092929     Owner:  NLM     Status:  MEDLINE    
Here we report on a 3.0 A crystal structure of a ternary complex of wild-type Thermus thermophilus argonaute bound to a 5'-phosphorylated 21-nucleotide guide DNA and a 20-nucleotide target RNA containing cleavage-preventing mismatches at the 10-11 step. The seed segment (positions 2 to 8) adopts an A-helical-like Watson-Crick paired duplex, with both ends of the guide strand anchored in the complex. An arginine, inserted between guide-strand bases 10 and 11 in the binary complex, locking it in an inactive conformation, is released on ternary complex formation. The nucleic-acid-binding channel between the PAZ- and PIWI-containing lobes of argonaute widens on formation of a more open ternary complex. The relationship of structure to function was established by determining cleavage activity of ternary complexes containing position-dependent base mismatch, bulge and 2'-O-methyl modifications. Consistent with the geometry of the ternary complex, bulges residing in the seed segments of the target, but not the guide strand, were better accommodated and their complexes were catalytically active.
Yanli Wang; Stefan Juranek; Haitao Li; Gang Sheng; Thomas Tuschl; Dinshaw J Patel
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Nature     Volume:  456     ISSN:  1476-4687     ISO Abbreviation:  Nature     Publication Date:  2008 Dec 
Date Detail:
Created Date:  2008-12-18     Completed Date:  2009-01-13     Revised Date:  2014-09-10    
Medline Journal Info:
Nlm Unique ID:  0410462     Medline TA:  Nature     Country:  England    
Other Details:
Languages:  eng     Pagination:  921-6     Citation Subset:  IM    
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MeSH Terms
Bacterial Proteins / chemistry*,  genetics,  metabolism*
Base Pair Mismatch
Base Pairing
Base Sequence
Crystallography, X-Ray
DNA / chemistry,  genetics,  metabolism*
Models, Molecular
Protein Conformation
RNA / chemistry,  genetics,  metabolism*
RNA Interference
RNA-Induced Silencing Complex / chemistry*,  genetics,  metabolism*
Substrate Specificity
Thermus thermophilus / chemistry*
Grant Support
Reg. No./Substance:
0/Bacterial Proteins; 0/RNA-Induced Silencing Complex; 63231-63-0/RNA; 9007-49-2/DNA

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